Micropathological patient replica based on unadulterated whole blood

Abstract

A patient replica is created from layered culture medium where solid culture medium is formed so that a discontinuity exists, for example, between layers of culture medium or between culture medium and the container holding the medium. An injection port is provided in registration with the discontinuity so that a sample of patient blood can be infused into the discontinuity to form a thin layer of blood in contact with a surface of culture medium. The thin layer of blood obviates the requirement for any anticoagulant allowing blood-borne pathogens to be readily cultured without using broth. Further antibiotics or other drug samples may be placed on an exposed surface of the culture medium above the blood layer so that the antibiotic can diffuse through the culture medium and reveal the sensitivities of the cultured pathogens. Other samples of pathogens or tissues can be placed on the surface of the culture medium so that effects of drugs or growth factors present in the patient blood can be observed thereby allowing the entrapped blood layer and culture medium, held to natural patient conditions, to act as a biological replica of the patient.

We claim:  
     
       1. An apparatus for culturing pathogens comprising: 
       a sterile container;  
       a layer of culture medium in the sterile container, the layer having a exposed surface in contact with the atmosphere in the container;  
       a discontinuity not in contact with the atmosphere in the container; and  
       means for injecting an aliquot of patient's blood into the discontinuity, forming a layer of patient blood not in contact with the atmosphere and leaving the exposed surface of the layer blood-free to accept a specimen.  
     
     
       2. The apparatus of claim  1 , wherein the sterile container is airtight and further contains a port in communication with the atmosphere in the container. 
     
     
       3. The apparatus of claim  2 , wherein the port functions as an airlock to permit addition or removal of gases. 
     
     
       4. The apparatus of claim  2 , wherein the port can be used for specimen insertion. 
     
     
       5. The apparatus of claim  1 , wherein a port is in communication with the discontinuity so that a partial vacuum can be drawn through said port while blood is being injected into the discontinuity. 
     
     
       6. The apparatus of claim  1 , wherein the sterile container has a top which contains a microbe air filter. 
     
     
       7. The apparatus of claim  1 , wherein the top of the sterile container has a plurality of injection ports. 
     
     
       8. The apparatus of claim  1 , wherein the discontinuity is formed by a layer of fabric. 
     
     
       9. The apparatus of claim  1 , wherein the discontinuity is formed by a layer of semipermeable membrane. 
     
     
       10. The apparatus of claim  8 , wherein the sterile container has an inner groove to accept the layer of fabric or to accept a ring holding said fabric. 
     
     
       11. The apparatus of claim  9 , wherein the sterile container has an inner groove to accept the layer of semipermeable membrane or to accept a ring holding said membrane. 
     
     
       12. The apparatus of claim  1  further comprising a second sterile container which encloses the sterile container. 
     
     
       13. The apparatus of claim  1 , wherein the means for injecting includes a tube to carry blood towards a central region of the discontinuity. 
     
     
       14. The apparatus of claim  13 , wherein the tube has, in a wall thereof, a plurality of apertures to facilitate spread of blood. 
     
     
       15. A process for culturing pathogens or cells in an environment duplicative of individual patient factors comprising the steps of: 
       providing a sterile culture dish containing a layer of culture medium, the layer having an exposed surface contacting the atmosphere within the culture dish, a discontinuity, and a means for injecting blood into the discontinuity;  
       injecting an aliquot of fresh unadulterated patient blood into the discontinuity so that a layer of blood is formed leaving said exposed surface blood-free; and  
       incubating the dish under conditions favorable to pathogen culture.  
     
     
       16. The process of claim  15 , wherein a sample of a patient's tissue is placed on the exposed surface of the layer of culture medium. 
     
     
       17. The process of claim  16 , wherein a sample of a patient's tissue is transported in a bag-in-a-bag device prior to use in the process. 
     
     
       18. The process of claim  15 , wherein cells from a patient's tissue are injected into the discontinuity. 
     
     
       19. The process of claim  15 , wherein a patient's specimen is placed on the exposed surface of the layer of culture medium so that materials diffusing from the blood layer can influence growth in the specimen. 
     
     
       20. The process of claim  15 , wherein a sample of a drug is placed on the exposed surface of the layer of culture medium so that material diffusing from the drug sample can influence growth. 
     
     
       21. The process of claim  15 , wherein a grid is placed on top of the exposed surface of the culture media to define and segment areas thereon. 
     
     
       22. The process of claim  15 , wherein a grid apparatus is placed on the exposed surface of the culture medium and samples of a drug are placed on the grid apparatus so that a material diffusing from the drug sample can influence growth in the blood layer. 
     
     
       23. The process of claim  22 , wherein the material diffusing from the drug sample passes through apertures a point projecting below the grid apparatus. 
     
     
       24. The process of claim  15 , wherein the container turned upside down to release said culture medium from the container, a bottom surface of said culture medium thereby providing a blood-free surface. 
     
     
       25. The process of claim  15 , wherein the steps of providing a sterile culture dish, injecting an aliquot and, incubating the dish are automated. 
     
     
       26. An improved device for culturing pathogens of the type where solid culture medium is placed in a sterile container and patient blood is added, the improvement comprising a discontinuity, not in contact with the atmosphere within the sterile container, and means for injecting a sample of patient's blood into the discontinuity to form a layer of blood whereby pathogens can be cultured without adding anticoagulants to the patient's blood. 
     
     
       27. The improved device of claim  26 , wherein the discontinuity is formed by a layer of fabric. 
     
     
       28. The improved device of claim  26 , wherein the discontinuity is formed by a layer of semipermeable membrane. 
     
     
       29. The improved device of claim  27 , wherein the fabric is held within a ring which fits within an inner groove of said improved device. 
     
     
       30. The improved device of claim  28 , wherein the membrane is held within a ring which fits within an inner groove of said improved device. 
     
     
       31. The improved device of claim  28  wherein two membranes are used forming a discontinuity between the layers of said membranes which accepts the infusion of blood and cells or serum. 
     
     
       32. The improved device of claim  31 , wherein the two membranes are held by a ring which fits within a groove on an inner surface of a side wall of said device, which sidewall has lids enclosing either end allowing access to both surfaces of the double membrane within. 
     
     
       33. A kit for culturing pathogens while minimizing exposure to medical personnel comprising: 
       a blood drawing set comprising:  
       a plurality of hollow needles attached to interconnected tubing;  
       valving means for controlling blood flow through the tubing;  
       a syringe for pulling or pushing blood through the tubing; and  
       a culturing device comprising:  
       a sterile container;  
       a layer of culture medium in the sterile container having a surface in contact with the atmosphere within the container;  
       a discontinuity not in contact with the atmosphere within the container; and  
       means for injecting an aliquot of patient's blood from the blood drawing set into the discontinuity, thereby forming a layer of patient blood wherein the blood from does not contact the atmosphere; and  
       means for disposing a patient specimen or inoculum on the surface of the layer in contact with the atmosphere.